Objective To evaluate the clinical application of automatic coagulation detection assembly line in high-throughput specimen detection.Methods The relevant information of sodium citrate anticoagulation samples in Zhongshan Hospital Affiliated to Fudan University from June to August 2021 was collected,inclu-ding sample collection time,receiving time,instrument sucking time,test completion time,and whether it pas-sed autoverification or not.The sample pretreatment time,testing time and turnaround time(TAT)of the au-tomatic coagulation detection assembly line were compared before and after installation,and the detection speed of the automatic coagulation detection assembly line was evaluated.Results The automatic coagulation detection line was expected to detect 650-900 samples per hour.The increase in the number of turbidimetric tests would slow down the detection speed of the instrument.Automatic coagulation detection assembly line test specimen to clinic and ward of pretreatment time and testing time were shorter than single detection,the differences were statistically significant(P＜0.05).The automatic coagulation detection assembly line could shorten TAT(P＜0.05).After the application of automatic coagulation detection assembly line,the autoveri-fication rate was 25.6％.Conclusion The automatic coagulation detection assembly line is suitable for high-throughput specimen detection in laboratory.Compared with stand-alone coagulation detection,the automatic coagulation detection assembly line could shorten TAT and testing time,and help to reduce the work pressure of laboratory personnel.

Objective:To investigate the characteristics and distribution of monoclonal gammopathy in clinical patients.Methods:A total of 936 171 patients (508 449 males and 427 722 females) who received capillary zone electrophoresis in Zhongshan Hospital Affiliated to Fudan University from January 2012 to December 2021 were selected, from which 14 945 patients with abnormal bands were screened as the study subjects, including 10 173 males and 4 772 females and the age 21-102 (65±13) years old. According to the age, patients were divided into 8 groups: 21-30 years old (168 cases), 31-40 years old (405 cases), 41-50 years old (1 326 cases), 51-60 years old (3 068 cases), 61-70 years old (4 985 cases), 71-80 years old (3 288 cases), 81-90 years old (1 519 cases), and≥91 years old (186 cases). The diagnostic results of the 14 945 patients with abnormal bands were collected and were divided into tumor group (5 196 cases) and non-tumor disease group (9 749 cases) according to the presence of tumor. The distribution of abnormal bands in different gender, age, and disease groups were retrospectively analyzed. Among the 14 945 patients, 4 988 cases underwent immunofixation electrophoresis, excluding 336 negative cases and 412 cases of double clonal bands reaction, and 4 240 patients with monoclonal immunoglobulin (M protein) reaction were selected as the study subjects, including 2 794 males and 1 446 females aged 21-102 (67±12) years old. They were divided into 8 groups according to the age: 21-30 years old (18 cases), 31-40 years old (91 cases), 41-50 years old (364 cases), 51-60 years old (862 cases), 61-70 years old (1 455 cases), 71-80 years old (904 cases), 81-90 years old (486 cases), and≥91 years old (60 cases). The diagnostic results and immunoglobin subtypes (IgA-κ, IgA-λ, IgG-κ, IgG-λ, IgM-κ, IgM-λ, κ, λ) of patients were collected, and the distribution of monoclonal gammopathy in different gender, age and disease groups were retrospectively analyzed.Results:Among 936 171 patients, 14 945 cases showed abnormal bands in electropherograms with a detection rate of 1.60%; the detection rates of abnormal bands in males and females were 2.00% (10 173/508 449) and 1.12% (4 772/427 722), respectively, with a statistically significant difference ( P<0.01). There was a significant difference in the detection rate of abnormal bands among different age groups ( P<0.01); among them, the highest detection rate of abnormal band in group of ≥91 years old was 5.98%, and the ratio of male to female was 1.67∶1. Among the 14 945 cases of abnormal bands, patients aged 51-60, 61-70 and 71-80 accounted for 20.53% (3 068 cases), 33.36% (4 985 cases) and 22.00% (3 288 cases), respectively, and the differences among the age groups were statistically significant (χ 2=115.82, P<0.01). In the tumor group, the top 3 tumors with abnormal bands were plasmacytoma with 1 123 cases, lymphoma with 289 cases, and leukemia with 49 cases. The detection rate of abnormal bands in electropherograms of plasmacytoma was 89.92% (1 123/1 249), which was higher than that in lymphoma and leukemia [6.73% (289/4 296) and 6.40% (49/766), respectively, P<0.01]. Among 4 240 patients with positive M protein, the proportion of 51-60, 61-70 and 71-80 years old patients were 20.33% (862/4 240), 34.32% (1 455/4 240) and 21.32% (904/4 240), respectively, and the differences among age groups were statistically significant ( P<0.01). The results of M protein types showed that the proportion of IgG-κ type was the highest in both genders, with 32.28% (902/2 794) in males and 34.30% (496/1 446) in females. In the 21-30, 31-40, and 41-50 age groups, the proportion of IgG-λ was the highest, which were 38.89% (7/18), 36.26% (33/91) and 34.07% (124/364) in these groups respectively. However, the proportions of IgG-κ were the highest in either of the 51-60, 61-70, 71-80, 81-90 and ≥91 years old groups, which were 33.87% (292/862), 34.16% (497/1 455), 31.53% (285/904), 34.57% (168/486), 28.33% (17/60), respectively, and the differences among all age groups and gender groups had statistical significance ( P<0.01). Among patients with positive M protein in the tumor group, plasmacytoma accounted for 14.22% (603/4 240), followed by lymphoma 6.30% (267/4 240); among non-tumor diseases, M proteinemia accounted for the highest proportion (7.24%, 307/4 240), followed by pulmonary infection (5.47%, 232/4 240). Conclusions:The detection rate of abnormal bands in capillary zone electrophoresis may increase with age, and is higher in males than in females in the same age group; different malignant tumor diseases can also show abnormal bands in capillary zone electrophoresis, but they are still mainly hematological tumors. Among the positive results of M protein, 61-70 years old group accounts for the highest proportion; the most common type of monoclonal gammopathy is IgG type; in the age group of 21-50 years, the proportion of IgG-λ type is the highest; in the group of >50 years old, the proportion of IgG-κ type is the highest; in the diagnosis of positive monoclonal gammopathy, the top 3 diseases are all hematological diseases, including plasmacytoma, monoclonal gammopathies and lymphoma.

Objectives:To observe the clinical characteristics of familial chylomicronemia syndrome (FCS) in Chinese population and explore the suitable diagnostic criteria of FCS in China.Methods:We screened 6856 patients with triglyceride ≥10 mmol/L from 9 hospitals in China between January 2010 and December 2023.The overall clinical information was collected and FCS-related gene testing was performed in 102 patients with high suspicion of FCS.Demographic characteristics were analyzed and FCS diagnosis was drafted.The FDA FCS diagnostic criteria was used to verify the FCS patients diagnosed in this study.At the same time,the prevalence of FCS patients with fasting triglyceride ≥10 mmol/L was explored in some of the above hospitals.Results:In this study,the diagnostic criteria for FCS in Chinese population were drafted based on European and American diagnostic criteria and Chinese clinical practice:(1) Fasting triglyceride ≥ 10 mmol/L after standard lipid-lowering treatment;(2) With one of the below-listed conditions:positive detection of FCS related genes;family history of hypertriglyceridemia pancreatitis (HTGP);history of pancreatitis in adolescence or adult HTGP;history of repeated hospitalization with unexplained abdominal pain.According to this criteria,60 were preliminarily diagnosed with FCS from the 102 patients enrolled.Their average age was (43.0±8.6) years old,male accounted for 70％,average triglyceride was (20.0±15.0) mmol/L.FCS related gene mutations were detected in 6 patients,all were lipoprotein lipase gene mutations.According to the American FCS diagnostic criteria,among the 60 FCS patients diagnosed in this study,the proportion of patients with "possible FCS" was 98.3％ (59/60),and the proportion of patients with "confirmed FCS" was 70.0％ (42/60).According to the diagnostic criteria of FCS patients in this study,the prevalence of FCS patients with TG ≥10 mmol/L is about 0.5％ (33/6722).Conclusions:The clinical and genetic characteristics of Chinese patients with FCS are consistent with those of European and American patients.The diagnostic criteria for FCS drafted in this study can be further verified and promoted in Chinese population.

Objectives:To observe the clinical characteristics of familial chylomicronemia syndrome (FCS) in Chinese population and explore the suitable diagnostic criteria of FCS in China.Methods:We screened 6856 patients with triglyceride ≥10 mmol/L from 9 hospitals in China between January 2010 and December 2023.The overall clinical information was collected and FCS-related gene testing was performed in 102 patients with high suspicion of FCS.Demographic characteristics were analyzed and FCS diagnosis was drafted.The FDA FCS diagnostic criteria was used to verify the FCS patients diagnosed in this study.At the same time,the prevalence of FCS patients with fasting triglyceride ≥10 mmol/L was explored in some of the above hospitals.Results:In this study,the diagnostic criteria for FCS in Chinese population were drafted based on European and American diagnostic criteria and Chinese clinical practice:(1) Fasting triglyceride ≥ 10 mmol/L after standard lipid-lowering treatment;(2) With one of the below-listed conditions:positive detection of FCS related genes;family history of hypertriglyceridemia pancreatitis (HTGP);history of pancreatitis in adolescence or adult HTGP;history of repeated hospitalization with unexplained abdominal pain.According to this criteria,60 were preliminarily diagnosed with FCS from the 102 patients enrolled.Their average age was (43.0±8.6) years old,male accounted for 70％,average triglyceride was (20.0±15.0) mmol/L.FCS related gene mutations were detected in 6 patients,all were lipoprotein lipase gene mutations.According to the American FCS diagnostic criteria,among the 60 FCS patients diagnosed in this study,the proportion of patients with "possible FCS" was 98.3％ (59/60),and the proportion of patients with "confirmed FCS" was 70.0％ (42/60).According to the diagnostic criteria of FCS patients in this study,the prevalence of FCS patients with TG ≥10 mmol/L is about 0.5％ (33/6722).Conclusions:The clinical and genetic characteristics of Chinese patients with FCS are consistent with those of European and American patients.The diagnostic criteria for FCS drafted in this study can be further verified and promoted in Chinese population.

Receptors, Purinergic ; Signal Transduction/physiology*

By the end of 2021, a total of 915000 cases of occupational pneumoconiosis and 450000 existing cases have been reported nationwide. Silicosis is a common and serious pneumoconiosis disease caused by long-term inhalation of large amounts of free silica dust and extensive nodular fibrosis in the lungs. Because its specific pathogenic mechanism has not been elucidated and the relevant research progress is slow, there is still a lack of effective therapeutic and interventional drugs. With the increase of national attention and the unique advantages of Chinese materia medica in the treatment of silicosis, more and more studies have been conducted on the treatment of silicosis with active ingredients of Chinese materia medica in China, but most of them are still in preclinical research stage. This article mainly introduced the pharmacological action and mechanism of selected active components of Chinese materia medica in the intervention of silicosis from three aspects: anti-inflammation, anti-oxidation, and intervention of apoptosis, providing ideas for subsequent research and development of new drugs for silicosis. This article argues, it is considered that some traditional Chinese medicines must observe the pathological changes in the treatment of silicosis in the overall animal experiment, clarify their pharmacodynamic effects, and further study the multiple targets and pathways involved in them to elucidate their specific mechanisms of action. At the same time, it can strengthen the analysis of active ingredients of traditional Chinese medicine, or modify the structure of active ingredients, and then enhance its pharmacological activity in the treatment of silicosis, realizing the transformation of preclinical research stage to the results of clinical research.

The global COVID-19 coronavirus pandemic has infected over 109 million people, leading to over 2 million deaths up to date and still lacking of effective drugs for patient treatment. Here, we screened about 1.8 million small molecules against the main protease (M^pro) and papain like protease (PL^pro), two major proteases in severe acute respiratory syndrome-coronavirus 2 genome, and identified 1851M^pro inhibitors and 205 PL^pro inhibitors with low nmol/l activity of the best hits. Among these inhibitors, eight small molecules showed dual inhibition effects on both M^pro and PL^pro, exhibiting potential as better candidates for COVID-19 treatment. The best inhibitors of each protease were tested in antiviral assay, with over 40% of M^pro inhibitors and over 20% of PL^pro inhibitors showing high potency in viral inhibition with low cytotoxicity. The X-ray crystal structure of SARS-CoV-2 M^pro in complex with its potent inhibitor 4a was determined at 1.8 Å resolution. Together with docking assays, our results provide a comprehensive resource for future research on anti-SARS-CoV-2 drug development.
Humans ; Antiviral Agents/chemistry* ; COVID-19 ; COVID-19 Drug Treatment ; High-Throughput Screening Assays ; Molecular Docking Simulation ; Protease Inhibitors/chemistry* ; SARS-CoV-2/enzymology* ; Viral Nonstructural Proteins

Objective:To investigate the expression of four cancer stem cell (CSC) markers (EpCAM, CD133, CD90 and CD24) in hepatocellular carcinoma tissues and peripheral blood circulating tumor cells (CTC),their value in the prognosis of patients with hepatocellular carcinoma.Methods:A total of 50 hepatocellular carcinoma tissues and 29 peripheral blood sample from 50 patients with hepatocellular cancer treated in Zhongshan Hospital Fudan University from October 2013 to September 2014 were collected and analyzed by flow cytometry or qRT-PCR to examine the expression of EpCAM, CD133, CD90 and CD24. The clinical data of patients were collected, including tumor size, tumor number, satellite lesions, vascular invasion, Edmondson stage, BCLC stage and liver cirrhosis, etc. The correlation between the expression of four markers in hepatocellular carcinoma tissues and CTC with the clinical data and survival time of patients were compared.Results:The positive expression rates of EpCAM, CD133, CD90 and CD24 in hepatocellular carcinoma tissues were 66% (33/50), 18% (9/50), 60% (30/50) and 56% (28/50); the positive expression rates in CTC were 55% (16/29), 38% (11/29), 31% (9/29) and 59% (17/29). CD90 expression in hepatocellular carcinoma tissue was positively correlated with the occurrence HCC liver cirrhosis ( P<0.05), while CD133 expression was negatively correlated with the 5-year survival rate of patients ( P<0.05). The expression of EpCAM and CD24 in peripheral blood CTC were closely related to the patient′s Edmondson stage ( P<0.05). The survival time of patients with CD133 positive expression in hepatocellular carcinoma tissue was lower than those without CD133 expression ( P<0.05); the survival rate of patients with EpCAM expressed in either tissue or peripheral blood CTC was lower than that of patients with EpCAM double negative expression ( P<0.05). The survival rate of patients with CD90 negative in HCC tissue and positive in peripheral blood was lower than that in patients with double negative/double positive in tissue and peripheral blood or patients positive in hepatocellular carcinoma tissue and negative in peripheral blood ( P<0.01). Conclusion:Different expression characteristics of four markers in cancer tissues and peripheral blood CTC might provide useful information about predicting prognosis of hepatocellular carcinoma. The expression of CD133 in tissues can be used as an important survival predictor of hepatocellular carcinoma patients. The differential expression of cancer markers in tissue samples and blood samples can provide more clinical prognostic information.

Objective:To establish autoverification rules for coagulation tests in multicenter cooperative units, in order to reduce workload for manual review of suspected results and shorten turnaround time (TAT) of test reports, while ensure the accuracy of results.Methods:A total of 14 394 blood samples were collected from fourteen hospitals during December 2019 to March 2020. These samples included: Rules Establishment Group 11 230 cases, including 1 182 cases for Delta check rules; Rules Validation Group 3 164 cases, including 487cases for Delta check; Clinical Application Trial Group 77 269 cases. Samples were analyzed for coagulation tests using Sysmex CS series automatic coagulation analyzers, and the clinical information, instrument parameters, test results, clinical diagnosis, medication history of anticoagulant and other relative results such as HCT, TG, TBIL, DBIL were summarized; on the basis of historical data, the 2.5 and 97.5 percentile of all data arranged from low to high were initially accumulated; on the basis of clinical suggestions, critical values and specific drug use as well as relative guidelines, autoverification rules and limits were established.The rules were then input into middleware, in which Stage I/Stage II validation was done. Positive coincidence, negative coincidence, false negative, false positive, autoverification pass rate, passing accuracy (coincidence of autoverification and manual verification) were calculated. Autoverification rules underwent trial application in coagulation results reports.Results:(1) The autoverification algorisms involve 33 rules regarding PT/INR, APTT, FBG, D-dimer, FDP,Delta check, reaction curve and sample abnormalities; (2)Autoverification Establishment Group showed autoverification pass rate was 68.42% (7 684/11 230), the false negative rate was 0%(0/11230), coincidence of autoverification and manual verification was 98.51%(11 063/11 230), in which positive coincidence and negative coincidence were respectively 30.09% (3 379/11 230) and 68.42%(7 684/11 230); Autoverification Validation Group showed autoverification pass rate was 60.37%(1 910/3 164), the false negative rate was 0%(0/11 230), coincidence of autoverification and manual verification was 97.79%(3 094/3 164), in which positive coincidence and negative coincidence were respectively 37.42%(1 184/3 164) and 60.37%(1 910/3 164); (3) Trialed implementation of these autoverification rules on 77 269 coagulation samples showed that the average TAT shortened by 8.5 min-83.1 min.Conclusions:This study established 33 autoverification rules in coagulation tests. Validation showedthese rules could ensure test quality while shortening TAT and lighten manual workload.